Short interfering RNAs targeting a vampire-bat related rabies virus phosphoprotein mRNA

Abstract

The aim of this study was to assess the in vitro and in vivo effects of short-interfering RNAs (siRNAs) against rabies virus phosphoprotein (P) mRNA in a post-infection treatment for rabies as an extension of a previous report (Braz J Microbiol. 2013 Nov 15;44(3):879–82). To this end, rabies virus strain RABV-4005 (related to the Desmodus rotundus vampire bat) were used to inoculate BHK-21 cells and mice, and the transfection with each of the siRNAs was made with Lipofectamine-2000™. In vitro results showed that siRNA 360 was able to inhibit the replication of strain RABV-4005 with a 1 log decrease in virus titter and 5.16-fold reduction in P mRNA, 24 h post-inoculation when compared to non-treated cells. In vivo, siRNA 360 was able to induce partial protection, but with no significant difference when compared to non-treated mice. These results indicate that, despite the need for improvement for in vivo applications, P mRNA might be a target for an RNAi-based treatment for rabies.

Rabies Prophylactic and Treatment Options: An In Vitro Study of siRNA- and Aptamer-Based Therapeutics

“…Unfortunately, the few studies undertaken for rabies have thus far demonstrated limited success [ 33 ]. The most recent study by Ono and colleagues (2017) demonstrated the capability of achieving an 80% reduction in viral titer in vitro with the siRNAs transfected using Lipofectamine 2000 but no statistically significant reduction was shown in vivo [ 34 ]. This demonstrated the feasibility of RABV inhibition using siRNAs, but highlighted the current shortcomings of effective, targeted delivery for successful in vivo application.…”

“…A recent study has provided the most promising results, with an siRNA (siRNA 360) producing an 80.6% reduction in RABV phosphoprotein (P) mRNA in in vitro trials when treatment occurred at 2 h post infection (p.i.) [ 34 ]. Yet, more evidence is required for siRNAs to be widely considered as an option in the pursuit of a RIG alternative or a potential treatment for rabies.…”

Evolution of Avian coronavirus (AvCoV) in BHK-21 and VERO cells

“…For absolute quantification, a plasmid with a 5'UTR sequence was used to construct a ten-fold dilution standard curve from 10 7 to 10 3 copies, which resulted in a linear equation (y=-3.357x + 38.05) with an R 2 value=1.0 and efficiency of 98.56%. The absolute values (copies/µL) were calculated with normalization using β-actin mRNA from cells, with GoTaq® Probe 1-Step RT-qPCR (Promega) performed according to the manufacturer's instructions and with primers (400 nM) and a probe (150 nM) described by Ono et al (2017).…”

Bioinformatics as a Tool to Identify Infectious Disease Pathogen Peptide Sequences as Targets for Antibody Engineering

“…Caprin-2 has a significant role in influencing phosphorylation of the Wnt-signaling pathways (PubMed:18,762,581) [27]. Caprin-2 also facilitates LRP6 phosphorylation by CDK14/CCNY during G2/M stage of the cell cycle, which may potentiate cells for transport or translation of mRNAs, modulate the expression of neuronal proteins involved in synaptic plasticity [28], while simultaneously influencing cell cycle signaling and regulation of viral transcription and replication [29,30]. leprae UniProtKB A0A197SEI9 (A0A197SEI9_MYCLR) (AA position: 170-2166) (Figure 16) 2′, 3′-cyclic-nucleotide 3′-phosphodiesterase is involved in RNA metabolism of the myelinating cell, CN37 (2′, 3′-cyclic-nucleotide 3′-phosphodiesterase) is the one of the most abundant myelin protein in nervous system.…”